Staff Publications

Staff Publications

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    'Staff publications' is the digital repository of Wageningen University & Research

    'Staff publications' contains references to publications authored by Wageningen University staff from 1976 onward.

    Publications authored by the staff of the Research Institutes are available from 1995 onwards.

    Full text documents are added when available. The database is updated daily and currently holds about 240,000 items, of which 72,000 in open access.

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Record number 348971
Title Selection and optimization of proteolytically stable llama single-domain antibody fragments for oral immunotherapy
Author(s) Harmsen, M.M.; Solt-Smits, C.B. van; Zijderveld-van Bemmel, A.M. van; Niewold, T.A.; Zijderveld, F.G. van
Source Applied Microbiology and Biotechnology 72 (2006)3. - ISSN 0175-7598 - p. 544 - 551.
DOI https://doi.org/10.1007/s00253-005-0300-7
Department(s) ASG Infectieziekten
CIDC - Divisie Bacteriologie en TSE's
ID - Dier en Omgeving
Publication type Refereed Article in a scientific journal
Publication year 2006
Keyword(s) infectious intestinal-diseases - chicken egg antibodies - in-vitro evolution - escherichia-coli - saccharomyces-cerevisiae - ribonuclease-a - stability - affinity - prophylaxis - expression
Abstract We previously demonstrated that oral application of the recombinant single-domain antibody fragment (VHH) clone K609, directed against Escherichia coli F4 fimbriae, reduced E. coli-induced diarrhoea in piglets, but only at high VHH doses. We have now shown that a large portion of the orally applied K609 VHH is proteolytically degraded in the stomach. Stringent selection for proteolytic stability identified seven VHHs with 7- to 138-fold increased stability after in vitro incubation in gastric fluid. By DNA shuffling we obtained four clones with a further 1.5- to 3-fold increased in vitro stability. These VHHs differed by at most ten amino acid residues from each other and K609 that were scattered over the VHH sequence and did not overlap with predicted protease cleavage sites. The most stable clone, K922, retained 41% activity after incubation in gastric fluid and 90% in jejunal fluid. Oral application of K922 to piglets confirmed its improved proteolytic stability. In addition, K922 bound to F4 fimbriae with higher affinity and inhibited fimbrial adhesion at lower VHH concentrations. K922 is thus a promising candidate for prevention of piglet diarrhoea. Furthermore, our findings could guide selection and improvement by genetic engineering of other recombinant antibody fragments for oral use.
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